Nonhuman Primate Lung Decellularization and Recellularization Using a Specialized Large-organ Bioreactor
Nonhuman Primate Lung Decellularization and Recellularization Using a Specialized Large-organ Bioreactor
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DOI：
10.3791/50825
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发表时间：
2013-12-01
影响因子：
1.2
通讯作者：
Bunnell, Bruce A.
中科院分区：
文献类型：
--
作者：
Bonvillain, Ryan W.;Scarritt, Michelle E.;Bunnell, Bruce A.
可获得的肺数量不足以满足当前和未来的器官移植需求。生物人工组织再生是传统器官移植的一个有吸引力的替代方案。这项技术利用器官的天然生物细胞外基质（ECM）作为支架，自体细胞或干细胞/祖细胞可以在支架上播种和培养，从而促进原始组织的再生。天然ECM是通过一个叫做脱细胞的过程分离出来的。脱细胞是通过用一系列洗涤剂、盐和酶处理组织来实现有效去除细胞物质，同时保持ECM完整。利用啮齿动物肺的脱细胞和随后的再细胞化进行的研究表明，在产生能够在体内进行气体交换的肺样组织方面取得了边际成功。虽然提供了基本的概念验证，但啮齿动物模型并不能直接转化为人类使用。非人灵长类动物（NHP）提供了一个更合适的模型，用于研究生物人工器官生产的最终临床应用。实现从NHP恒河猴肺获得完全脱细胞的方案提出。由此产生的无细胞肺可以用多种细胞播种，包括间充质干细胞和内皮细胞。该手稿还描述了一种生物反应器系统的发展，其中细胞种子猕猴肺可以在负压通气提供的机械拉伸和应变条件下培养，以及通过脉管系统进行脉动灌注；已知这些力分别沿着肺和内皮细胞谱系指导分化。给出了脱细胞和细胞播种的代表性结果。
There are an insufficient number of lungs available to meet current and future organ transplantation needs. Bioartificial tissue regeneration is an attractive alternative to classic organ transplantation. This technology utilizes an organ's natural biological extracellular matrix (ECM) as a scaffold onto which autologous or stem/progenitor cells may be seeded and cultured in such a way that facilitates regeneration of the original tissue. The natural ECM is isolated by a process called decellularization. Decellularization is accomplished by treating tissues with a series of detergents, salts, and enzymes to achieve effective removal of cellular material while leaving the ECM intact. Studies conducted utilizing decellularization and subsequent recellularization of rodent lungs demonstrated marginal success in generating pulmonary-like tissue which is capable of gas exchange in vivo. While offering essential proof-of-concept, rodent models are not directly translatable to human use. Nonhuman primates (NHP) offer a more suitable model in which to investigate the use of bioartificial organ production for eventual clinical use.The protocols for achieving complete decellularization of lungs acquired from the NHP rhesus macaque are presented. The resulting acellular lungs can be seeded with a variety of cells including mesenchymal stem cells and endothelial cells. The manuscript also describes the development of a bioreactor system in which cell-seeded macaque lungs can be cultured under conditions of mechanical stretch and strain provided by negative pressure ventilation as well as pulsatile perfusion through the vasculature; these forces are known to direct differentiation along pulmonary and endothelial lineages, respectively. Representative results of decellularization and cell seeding are provided.